Witches’ broom disease of lime (WBDL), associated with ‘Candidatus Phytoplasma aurantifolia,’ is a serious disease of acid lime in Southern Iran. In order to characterize ‘Ca. P. aurantifolia’ strains, a multilocus sequence analysis protocol was developed based on different genomic loci, including 16S rRNA, 16S-23S rRNA intergenic spacer (IS), ribosomal protein genes rpl22 and rps3, cell division protein, and group II intron reverse transcriptase/mature gene, in 46 phytoplasma strains. Analyses of virtual RFLP, phylogenetics, and DNA sequence identity of 16S rRNA of Iranian phytoplasma strains associated with symptomatic citrus plants indicated that the Iranian strains were classified in ribosomal subgroup 16SrII-B. In total, three genotypes were recognized based on 16Sr virtual RFLP profiles. Phylogenetic analysis based on the rp and ftsH gene sequences showed clustering of all the strains as one group. Comparative sequence and phylogenetic analysis of group II intron genes indicated the highest differentiation of ‘Ca. P. aurantifolia’–related strains in four genotypes. The polymorphism of group II intron reverse transcriptase/mature genes fragmentation provides finer differentiation of closely related ‘Ca P. aurantifolia’ strains and reveals higher genetic diversity than previously reported.

中文翻译：

---

柑橘属植物中与女巫扫帚病相关的'Candidatus Phytoplasma aurantifolia'的多位点基因分型。

与'Candidatus Phytoplasma aurantifolia'相关的女巫的石灰病（WBDL）是伊朗南部的一种严重的酸性石灰病。为了表征'Ca。P. aurantifolia' 菌株，基于不同基因组位点开发了多位点序列分析方案，包括 16S rRNA、16S-23S rRNA 基因间隔区 (IS)、核糖体蛋白基因 rpl22 和 rps3、细胞分裂蛋白和 II 组内含子逆转录酶/成熟基因，在 46 个植原体菌株中。对与有症状的柑橘植物相关的伊朗植原体菌株的 16S rRNA 的虚拟 RFLP、系统发育和 DNA 序列同一性的分析表明，伊朗菌株被归类为核糖体亚群 16SrII-B。根据 16Sr 虚拟 RFLP 配置文件，总共识别了三种基因型。基于 rp 和 ftsH 基因序列的系统发育分析显示所有菌株聚类为一组。II组内含子基因的比较序列和系统发育分析表明'Ca'的最高分化。P. aurantifolia' 相关菌株的四种基因型。II 组内含子逆转录酶/成熟基因片段的多态性为密切相关的'Ca P. aurantifolia' 菌株提供了更精细的分化，并显示出比以前报道的更高的遗传多样性。